The present invention is based, in part, on the discovery that aspirin reverses insulin resistance in liver and fat cells, e.g., by targeting IKK-xcex2. Thus, IKK-xcex2 was discovered as a target for identifying compounds for the treatment of disorders associated with insulin resistance.
Accordingly, in one aspect, the invention features a method of identifying, evaluating or making a compound or agent, e.g., a candidate compound or agent, for treatment of a disorder characterized by insulin resistance. The method includes evaluating the ability of a compound or agent to interact with, e.g., bind, IKK-xcex2, to thereby identify a compound or agent for the treatment of a disorder characterized by insulin resistance.
In a preferred embodiment, the disorder is diabetes, e.g., Type I or Type II diabetes, obesity, polycystic ovarian disease or syndrome X.
In a preferred embodiment, the compound is: a polypeptide, e.g., a randomly generated polypeptide which binds IKK-xcex2; an antibody, e.g., an intrabody or a randomly generated antibody which binds IKK-xcex2; a small molecule, e.g., a small molecule which binds IKK-xcex2.
In a preferred embodiment, the method further includes contacting the identified compound with IKK-xcex2, e.g., purified IKK-xcex2, to thereby evaluate binding between the compound and IKK-xcex2.
In a preferred embodiment, the method further includes contacting the identified compound with a cell, e.g., a fat cell or a liver cell, to thereby evaluate the effect of the compound on an IKK-xcex2 activity of the cell. For example, the ability of the compound to modulate, e.g., reduce or reverse, insulin resistance in a cell.
In a preferred embodiment, the method further includes administering the identified compound to a subject to, evaluate the effect of the compound on insulin resistance. In a preferred embodiment, the subject is a mouse (e.g., a NOD mouse, an ob/ob mouse, a db/db mouse) or a rat (e.g., a Zucker fatty rat, a streptozotocin rat).
In another aspect, the invention features a method of identifying a compound or agent for treatment of a disorder characterized by insulin resistance. The method includes contacting IKK-xcex2, or a cell expressing IKK-xcex2 with a test compound; and determining whether the test compound binds to IKK-xcex2, to thereby identify a compound.
Methods for identifying a compound or an agent can be performed, for example, using a cell free assay. For example, the IKK-xcex2 can be immobilized to a suitable substrate, e.g., glutathoine sepharose beads or glutathoine derivatized microtiter plates, using a fusion protein which allows for IKK-xcex2 to bind to the substrate, e.g., a glutathoine-S-transferase/IKK-xcex2 fusion protein.
In a preferred embodiment, the ability of a test compound to bind IKK-xcex2 can be determined by detecting the formation of a complex between IKK-xcex2 and the compound. The presence of the compound in complex indicates the ability to bind IKK-xcex2.
In a preferred embodiment, IKK-xcex2 is further contacted with aspirin.
In another preferred embodiment, a compound is identified using a cell based assay. These methods include identifying a compound based on its ability to modulate, e.g., inhibit, an IKK-xcex2 activity of the cell. For example, the ability of a compound to modulate, e.g., reduce or reverse, insulin resistance in a cell, e.g., a fat cell or a liver cell, can be determined.
In a preferred embodiment, the method further includes contacting the identified compound with IKK-xcex2, e.g., purified IKK-xcex2, to thereby evaluate binding between the compound and IKK-xcex2.
In a preferred embodiment, the method further includes contacting the identified compound with a cell, e.g., a fat cell or a liver cell, to thereby evaluate the effect of the compound on an IKK-xcex2 activity of the cell. For example, the ability of the compound to modulate, e.g., reduce or reverse, insulin resistance in a cell can be evaluated.
In a preferred embodiment, the method further includes administering the identified compound to a subject to evaluate the effect of the compound on insulin resistance. In a preferred embodiment, the subject is a mouse (e.g., a NOD mouse, an ob/ob mouse, a db/db mouse) or a rat (e.g., a Zucker fatty rat, a streptozotocin rat).
In a preferred embodiment, the compound is: a polypeptide, e.g., a randomly generated polypeptide which interacts with, e.g., binds, IKK-xcex2; an antibody, e.g., an intrabody or a randomly generated antibody which interacts with IKK-xcex2; a small molecule, e.g., a small molecule which interacts with IKK-xcex2.
In a preferred embodiment, the compound is a compound other than aspirin.
In a preferred embodiment, the disorder is diabetes, e.g., Type I or Type II diabetes, obesity, polycystic ovarian disease or syndrome X.
In another aspect, the invention features a method of identifying a compound or agent for treatment of diabetes, e.g., Type I or Type II diabetes. The method includes contacting IKK-xcex2, or a cell expressing IKK-xcex2 with a test compound; and determining whether the test compound binds to IKK-xcex2, to thereby identify a compound for treatment of diabetes.
Methods for identifying a compound or an agent can be performed, for example, using a cell free assay. For example, the IKK-xcex2 can be immobilized to a suitable substrate, e.g., glutathoine sepharose beads or glutathoine derivatized microtiter plates, using a fusion protein which allows for IKK-xcex2 to bind to the substrate, e.g., a glutathoine-S-transferase/IKK-xcex2 fusion protein.
In a preferred embodiment, the ability of a test compound to bind IKK-xcex2 can be determined by detecting the formation of a complex between IKK-xcex2 and the compound. The presence of the compound in complex indicates the ability to bind IKK-xcex2.
In a preferred embodiment, IKK-xcex2 is further contacted with aspirin.
In another preferred embodiment, a compound is identified using a cell based assay. These methods include identifying a compound based on its ability to modulate, e.g., inhibit, an IKK-xcex2 activity of the cell. For example, the ability of a compound to modulate, e.g., reduce or reverse, insulin resistance in a cell, e.g., a fat cell or a liver cell, can be determined.
In a preferred embodiment, the method further includes contacting the identified compound with IKK-xcex2, e.g., purified IKK-xcex2, to thereby evaluate binding between the compound and IKK-xcex2.
In a preferred embodiment, the method further includes contacting the identified compound with a cell, e.g., a fat cell or a liver cell, to thereby evaluate the effect of the compound on an IKK-xcex2 activity of the cell. For example, the ability of the compound to modulate, e.g., reduce or reverse, insulin resistance in a cell.
In a preferred embodiment, the method further includes administering the identified compound to a subject to evaluate the effect of the compound on insulin resistance. In a preferred embodiment, the subject is a mouse (e.g., a NOD mouse, an ob/ob mouse, a db/db mouse) or a rat (e.g., a Zucker fatty rat, a streptozotocin rat).
In a preferred embodiment, the compound is: a polypeptide, e.g., a randomly generated polypeptide which interacts with, e.g., binds, IKK-xcex2; an antibody, e.g., an intrabody or a randomly generated antibody which interacts with IKK-xcex2; a small molecule, e.g., a small molecule which interacts with IKK-xcex2.
In a preferred embodiment, the compound is a compound other than aspirin.
In another aspect, the invention features a method of treating a subject having a disorder characterized by insulin resistance. The method includes: administering a compound or agent which interacts with, e.g., binds, IKK-xcex2, to thereby treat the disorder.
In a preferred embodiment, the disorder is diabetes, e.g., Type I or Type II diabetes, obesity, polycystic ovarian disease or syndrome X.
In a preferred embodiment, the compound is: a compound other than aspirin; a polypeptide, e.g., a randomly generated polypeptide which interacts with IKK-xcex2; an antibody, e.g., an intrabody or a randomly generated antibody which interacts with IKK-xcex2; a small molecule, e.g., a small molecule which interacts with IKK-xcex2. In a preferred embodiment, the method includes administering a nucleic acid encoding one of the above-described compounds. In a preferred embodiment, the compound is a compound identified by a method described herein.
In a preferred embodiment, the compound is administered parenterally, e.g., intravenously, intradermally, subcutaneously, orally (e.g., inhalation). In a preferred embodiment, the administration of the compound is time-released.
In a preferred embodiment, the subject is a human. In another preferred embodiment, the subject is a NOD mouse, an ob/ob mouse, a db/db mouse, a Zucker fatty rat, or a streptozotocin induced rat.
In another aspect, the invention features a method of treating a subject having diabetes, e.g., Type I or Type II diabetes. The method includes administering to a subject a compound or agent which interacts with, e.g., binds, IKK-xcex2, to thereby treat the diabetes.
In a preferred embodiment, the compound is: a compound other than aspirin; a polypeptide, e.g., a randomly generated polypeptide which interacts with IKK-xcex2; an antibody, e.g., an intrabody, e.g., an anti-IKK-xcex2 antibody or a randomly generated antibody which interacts with IKK-xcex2, a small molecule, e.g., a small molecule which interacts with IKK-xcex2. In a preferred embodiment, the method includes administering a nucleic acid encoding one of the above-described compounds. In a preferred embodiment, the compound is a compound identified by a method described herein.
In a preferred embodiment, the compound is administered parenterally, e.g., intravenously, intradermally, subcutaneously, orally (e.g., inhalation). In a preferred embodiment, the administration of the compound is time-released.
In a preferred embodiment, the subject is a human. In another preferred embodiment, the subject is a NOD mouse, an ob/ob mouse, a db/db mouse, a Zucker fatty rat, or a streptozotocin induced rat.
In another aspect, the invention features compounds for the treatment of disorders characterized by insulin resistance, identified by the methods described herein.
The terms protein, polypeptide and peptide are used interchangeably herein.
A subject, as used herein, refers to a mammal, e.g., a human. It can also refer to an experimental animal, e.g., an animal model for an insulin-related disorder, e.g., a NOD mouse, an ob/ob mouse, a db/db mouse, a Zucker fatty rat, or a streptozotocin induced mouse or rat. The subject can be a human which is at risk for a disorder characterized by insulin resistance. Such disorders include diabetes, e.g., Type I or Type II, obesity, polycystic ovarian disease and syndrome X.